Volume 36 Issue 3
May  2015
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Lin ZHANG, Xu-He HUANG, Ping-Ping ZHOU, Guo-Long YU, Jin YAN, Bing QIN, Xin-Ge YAN, Li-Mei DIAO, Peng LIN, Yi-Qun KUANG. Establishment of HIV-1 model cell line GHOST(3) with stable DRiP78 and NHERF1 knockdown. Zoological Research, 2015, 36(3): 161-166.
Citation: Lin ZHANG, Xu-He HUANG, Ping-Ping ZHOU, Guo-Long YU, Jin YAN, Bing QIN, Xin-Ge YAN, Li-Mei DIAO, Peng LIN, Yi-Qun KUANG. Establishment of HIV-1 model cell line GHOST(3) with stable DRiP78 and NHERF1 knockdown. Zoological Research, 2015, 36(3): 161-166.

Establishment of HIV-1 model cell line GHOST(3) with stable DRiP78 and NHERF1 knockdown

Funds:  This study was supported by the Science and Technology Planning Project of Guangdong Province of China (2012B031800267); the Natural Science Foundation of Guangdong Province of China (S2013010011860) and the National Natural Science Foundation of China (31200130 and 81371812)
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  • Corresponding author: Yi-Qun KUANG
  • Received Date: 2015-01-21
  • Rev Recd Date: 2015-03-20
  • Publish Date: 2015-05-08
  • Chemokine receptors CXCR4 and CCR5 are indispensable co-receptors for HIV-1 entry into host cells. In our previous study, we identified that dopamine receptor-interacting protein 78 (DRiP78) and Na+-H+ exchanger regulatory factor 1 (NHERF1) are the CXCR4 and CCR5 homo- or hetero-dimer-interacting proteins. DRiP78 and NHERF1 are able to influence the co-receptor internalization and intracellular trafficking. Over-expression of NHERF1 affects the ligands or HIV-1 gp120-induced CCR5 internalization and HIV-1 production. It is reasonable to speculate that DRiP78 and NHERF1, as well as the signaling pathways involved in viral replication, would probably affect HIV-1 replication through regulating the co-receptors. In this present study, we designed two short hairpin RNAs (shRNAs) targeting the DRiP78 and NHERF1, respectively, and constructed the pLenti6/BLOCK-iT-DEST lentiviral plasmids expressing DRiP78 or NHERF1 shRNA. The packaged lentiviruses were used to transduce the widely-applied HIV-1 model cell line GHOST(3). Then, cells with stable knockdown were established through selecting transduced cells with Blasticidin. This study, for the first time, reported the establishment of the GHOST(3) with DRiP78 and NHERF1 knockdown, which is the first stable cell line with HIV-1 co-receptor-interacting molecular defects.
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  • [1]
    Bermak JC, Li M, Bullock C, Zhou QY. 2001. Regulation of transport of the dopamine D1 receptor by a new membrane-associated ER protein. Nature Cell Biology, 3(5): 492-498.
    Cao TT, Deacon HW, Reczek D, Bretscher A, von Zastrow M. 1999. A kinase-regulated PDZ-domain interaction controls endocytic sorting of the β2-adrenergic receptor. Nature, 401(6750): 286-290.
    Cicala C, Arthos J, Martinelli E, Censoplano N, Cruz CC, Chung E, Selig SM, Van Ryk D, Yang J, Jagannatha S, Chun TW, Ren P, Lempicki RA, Fauci AS. 2006. R5 and X4 HIV envelopes induce distinct gene expression profiles in primary peripheral blood mononuclear cells. Proceedings of the National Academy of Sciences of the United States of America, 103(10): 3746-3751.
    Dalgleish AG, Beverley PC, Clapham PR, Crawford DH, Greaves MF, Weiss RA. 1984. The CD4 (T4) antigen is an essential component of the receptor for the AIDS retrovirus. Nature, 312(5996): 763-767.
    Deng H, Liu R, Ellmeier W, Choe S, Unutmaz D, Burkhart M, Di Marzio P, Marmon S, Sutton RE, Hill CM, Davis CB, Peiper SC, Schall TJ, Littman DR, Landau NR. 1996. Identification of a major co-receptor for primary isolates of HIV-1. Nature, 381(6584): 661-666.
    Dupré DJ, Robitaille M, Richer M, Ethier N, Mamarbachi AM, Hébert TE. 2007. Dopamine receptor-interacting protein 78 acts as a molecular chaperone for Gγ subunits before assembly with Gβ. Journal of Biological Chemistry, 282(18): 13703-13715.
    Feng Y, Broder CC, Kennedy PE, Berger EA. 1996. HIV-1 entry cofactor: functional cDNA cloning of a seven-transmembrane, G protein-coupled receptor. Science, 272(5263): 872-877.
    Hammad MM, Kuang YQ, Yan R, Allen H, Dupré DJ. 2010. Na+/H+ exchanger regulatory factor-1 is involved in chemokine receptor homodimer CCR5 internalization and signal transduction but does not affect CXCR4 homodimer or CXCR4-CCR5 heterodimer. Journal of Biological Chemistry, 285(45): 34653-34664.
    Hung AY, Sheng M. 2002. PDZ domains: structural modules for protein complex assembly. Journal of Biological Chemistry, 277(8): 5699-5702.
    Kelley WL. 1998. The J-domain family and the recruitment of chaperone power. Trends in Biochemical Sciences, 23(6): 222-227.
    Klatzmann D, Champagne E, Chamaret S, Gruest J, Guetard D, Hercend T, Gluckman JC, Montagnier L. 1984. T-lymphocyte T4 molecule behaves as the receptor for human retrovirus LAV. Nature, 312(5996): 767-768.
    Kuang YQ, Charette N, Frazer J, Holland PJ, Attwood KM, Dellaire G, Dupré DJ. 2012a. Dopamine receptor-interacting protein 78 acts as a molecular chaperone for CCR5 chemokine receptor signaling complex organization. PLoS One, 7(7): e40522.
    Kuang YQ, Pang W, Zheng YT, Dupré DJ. 2012b. NHERF1 regulates gp120-induced internalization and signaling by CCR5, and HIV-1 production. European Journal of Immunology, 42(2): 299-310.
    Lazar CS, Cresson CM, Lauffenburger DA, Gill GN. 2004. The Na+/H+ exchanger regulatory factor stabilizes epidermal growth factor receptors at the cell surface. Molecular Biology of the Cell, 15(12): 5470-5480.
    Leclerc PC, Auger-Messier M, Lanctot PM, Escher E, Leduc R, Guillemette G. 2002. A polyaromatic caveolin-binding-like motif in the cytoplasmic tail of the type 1 receptor for angiotensin II plays an important role in receptor trafficking and signaling. Endocrinology, 143(12): 4702-4710.
    Li JG, Chen C, Liu-Chen LY. 2002. Ezrin-radixin-moesin-binding phosphoprotein-50/Na+/H+ exchanger regulatory factor (EBP50/NHERF) blocks U50, 488H-induced down-regulation of the human κ opioid receptor by enhancing its recycling rate. Journal of Biological Chemistry, 277(30): 27545-27552.
    Málaga-Diéguez L, Yang Q, Bauer J, Pankevych H, Freissmuth M, Nanoff C. 2010. Pharmacochaperoning of the A1 adenosine receptor is contingent on the endoplasmic reticulum. Molecular Pharmacology, 77(6): 940-952.
    Mörner A, Björndal A, Albert J, Kewalramani VN, Littman DR, Inoue R, Thorstensson R, Fenyö EM, Björling E. 1999. Primary human immunodeficiency virus type 2 (HIV-2) isolates, like HIV-1 isolates, frequently use CCR5 but show promiscuity in coreceptor usage. Journal of Virology, 73(3): 2343-2349.
    Tsibris AM, Kuritzkes DR. 2007. Chemokine antagonists as therapeutics: focus on HIV-1. Annual Review of Medicine, 58: 445-459.
    Wang B, Bisello A, Yang Y, Romero GG, Friedman PA. 2007. NHERF1 regulates parathyroid hormone receptor membrane retention without affecting recycling. Journal of Biological Chemistry, 282(50): 36214-36222.
    Weinman EJ, Steplock D, Wang Y, Shenolikar S. 1995. Characterization of a protein cofactor that mediates protein kinase A regulation of the renal brush border membrane Na+-H+ exchanger. The Journal of Clinical Investigation, 95(5): 2143-2149.
    Yi Z, Sperzel L, Nürnberger C, Bredenbeek PJ, Lubick KJ, Best SM, Stoyanov CT, Law LM, Yuan Z, Rice CM, MacDonald MR. 2011. Identification and characterization of the host protein DNAJC14 as a broadly active flavivirus replication modulator. PLoS Pathogens, 7(1): e1001255.
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